EP0116251A1 - Verfahren zum Depolymerisieren und Sulfatieren von Polysacchariden - Google Patents

Verfahren zum Depolymerisieren und Sulfatieren von Polysacchariden Download PDF

Info

Publication number
EP0116251A1
EP0116251A1 EP83402534A EP83402534A EP0116251A1 EP 0116251 A1 EP0116251 A1 EP 0116251A1 EP 83402534 A EP83402534 A EP 83402534A EP 83402534 A EP83402534 A EP 83402534A EP 0116251 A1 EP0116251 A1 EP 0116251A1
Authority
EP
European Patent Office
Prior art keywords
heparin
depolymerized
mixture
sulfation
sulfuric acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP83402534A
Other languages
English (en)
French (fr)
Other versions
EP0116251B1 (de
Inventor
Annamaria Naggi
Giangiacomo Torri
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sanofi SA
Original Assignee
Sanofi SA
Anic SpA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from FR8221934A external-priority patent/FR2538404B1/fr
Priority claimed from FR8319506A external-priority patent/FR2555993B1/fr
Application filed by Sanofi SA, Anic SpA filed Critical Sanofi SA
Priority to AT83402534T priority Critical patent/ATE31419T1/de
Publication of EP0116251A1 publication Critical patent/EP0116251A1/de
Application granted granted Critical
Publication of EP0116251B1 publication Critical patent/EP0116251B1/de
Expired legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof

Definitions

  • the present invention relates to a process for the depolymerization and sulfation of polysaccharides.
  • Sulphated polysaccharides are compounds of great importance in the cosmetic, textile, food and pharmaceutical industries. In particular, their use is recommended in the prevention of venous thrombosis (I.B. Jacques, Pharmacological Reviews 1979, 31, 99-166).
  • Low molecular weight sulfated polysaccharides are obtained by sulfating low molecular weight polysaccharides. Sulfation is generally carried out by treatment with chlorosulfonic acid in pyridine (KL Wolfrom et al., J. Am. Chem. Soc., 1953, 75, 1519) or else with adducts of trioxide of sulfur (sulfuric anhydride) with aprotic solvents (RL Whilster, WW Spencer, Methods Carbohydrate Chem., 1964, 4, 297-298; RL Whilster, ibid., 1972, 6, 426-429).
  • Low molecular weight polysaccharides are generally obtained by fractionation of a set of species with varied molecular weight or else by controlled depolymerization of unfractionated polysaccharides using nitrous acid.
  • the depolymerization processes also include hydrolysis of said N-sulfated groups, essential for the biological activity of heparin.
  • the subject of the present invention is a process for the depolymerization and the sulfation of polysaccharides, characterized in that a polysaccharide is treated with a mixture of sulfuric acid and chlorosulfonic acid.
  • the two acids in the mixture are concentrated and, preferably, their concentration is at least greater than 95% by weight.
  • the ratio of the two acids is very variable and can range from traces of chlorosulfonic acid in sulfuric acid to a sulfuric acid: chlorosulfonic acid ratio 1: 4 by volume.
  • the sulfuric acid: chlorosulfonic acid ratio varies from 4: 1 to 1: 1, with a ratio of about 2: 1 being particularly preferred.
  • the reaction temperature and the concentration of starting material in the sulfuric acid / chlorosulfonic acid mixture can vary depending on the nature of the substrate.
  • the low solubility of cellulose suggests higher dilutions, while in the case of chitosan, it is possible to use a higher concentration and to carry out the reaction at relatively low temperature.
  • the reaction temperature can vary between -20 and + 40 ° C; after a variable period due to the temperature, from a few minutes to 2 hours, the reaction is completed and the depolymerized and sulfated polysaccharide is isolated according to conventional techniques, for example by neutralization and dialysis, by chromatography or by lyophilization.
  • the depolymerized and sulfated polysaccharide can also be isolated by pouring the reaction mixture into a solvent in which the final product is insoluble, for example an apotic aprotic solvent, such as diethyl ether, by filtering the precipitate which has formed and by purifying it according to techniques known in sugar chemistry.
  • a solvent in which the final product is insoluble for example an apotic aprotic solvent, such as diethyl ether
  • the depolymerized and sulfated polysaccharides can be isolated in the form of alkaline salts according to the usual techniques, for example by lyophilization or by evaporation under pressure. reduced, and characterized by known physico-chemical methods.
  • salts for example the calcium salt
  • the alkaline salts preferably the sodium salt
  • the appropriate salt for example a calcium salt, optionally using a resin. ion exchange.
  • starting polysaccharides having a very high degree of polymerization for example in the case of chitosan, chitin or cellulose
  • the product thus partially depolymerized beforehand can be subsequently depolymerized and sulfated according to the process of the present invention.
  • the starting polysaccharide having a very high molecular weight can also be subjected twice to the process of the present invention. In this case, it is not even necessary to isolate the depolymerized product; a second quantity of the sulfuric acid / chlorosulfonic acid mixture can be added after, for example, the first hour of reaction.
  • This procedure does not involve any degradation or subsequent sulfation.
  • a depolymerized compound selectively sulfated in position 6, namely on the primary hydroxyl is obtained by this procedure.
  • Suitable starting materials are heparin, heparansulfates, chitosan, chitin, cellulose, starch, guaran, chondroitinsulfates, polyxylans, inulin, dermatansulfate, keratan, mannan, scleroglucan , galactomannans, dextrans, galactans, xantane.
  • the advantage of the process of the present invention lies in its selectivity and in its workability.
  • heparin for example, a depolymerized and supersulfated heparin is obtained having a molecular weight between 2000 and 9000 and having a higher degree of sulfation than that of the starting heparin.
  • this depolymerized and "supersulfated" heparin all of the primary hydroxyl groups are sulfated.
  • chitosan by reaction with a sulfuric acid / chlorosulfonic acid mixture according to the present invention, a chitosan is obtained of which the degree of depolymerization is not known because its molecular weight, like that of the starting compound, is too high, but that is assumed to be depolymerized.
  • This compound has selectively sulfated primary hydroxyls, with no variation in the secondary hydroxyl and the free amino group.
  • Cellulose, starch, chitin behave like chitosan.
  • the degree of depolymerization varies according to the molecular weight of the starting material and the stability of the glucose linkages. From this point of view, chitin and chitosan are more stable and depolymerization is minor.
  • Chondroitinsulfate and dermatansulfate are less stable and depolymerization can reach tri- and tetrasaccharides.
  • the degree of depolymerization can be controlled by suitably changing the sulfuric acid / chlorosulfonic acid ratio, the reaction time as well as the concentration of the starting material in the mixture of the two acids.
  • the mixture is left for one hour at room temperature, then it is poured into 250 ml of cold diethyl ether (from -10 to + 4 ° C.), it is filtered, the precipitate obtained is dissolved in water, neutralizes the solution thus obtained with 0.5 M sodium hydroxide and dialyzes against distilled water in 3500 D membranes (THOMAS DIALYZER TUBING 3787-H47) having a diameter of 11 mm in order to remove the salts and smaller reaction products.
  • the product thus obtained is dissolved in an aqueous 0.1 M calcium chloride solution, 0.5 M calcium hydroxide is added to the solution thus obtained up to pH 8 and dialyzed first against 500 ml 0.1 M calcium chloride solution and then against distilled water. By slow evaporation under reduced pressure, the calcium salt of a depolymerized and supersulfated heparin is obtained in the form of a white powder.
  • heparin from pig intestinal mucosa 500 mg are added (PROQUIFIN, batch 7926-7935, code number: D-212) having a degree of sulfation of 1.95 and a molecular weight of 13500.
  • a depolymerized heparin is obtained with a yield of 98% and supersulfated (code number: AH-104).
  • heparin from pig intestinal mucosa 500 mg are added (TEROPMON, batch 575/018, code number: D-98) having a molecular weight of 13500 and a degree of sulfation of 1.8.
  • FIG. 9 also shows that the compound AH-118 has a photodensitometric profile similar to those of the compounds AH-16 (example 1, FIG. 1) and AH-17 (example 4, FIG. 6), while the starting heparin D -98 shows itself to be very heterogeneous and quite different from the starting heparins used in examples 1 and 4.
  • - Electrophoresis in barium acetate shows that AH-118 has an electrophoretic characteristic "slow-moving", at the difference from the starting heparin D-98 which contains both "slow-moving" species and "fast-moving" species.
  • Figure 10 also confirms the indications of Figure 9 and, in addition, surprisingly shows that the compound AH-118 is not significantly different from the AH-108 of Example 11, although the heparins are very different.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Materials Engineering (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
EP83402534A 1982-12-28 1983-12-26 Verfahren zum Depolymerisieren und Sulfatieren von Polysacchariden Expired EP0116251B1 (de)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AT83402534T ATE31419T1 (de) 1982-12-28 1983-12-26 Verfahren zum depolymerisieren und sulfatieren von polysacchariden.

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
FR8221934 1982-12-28
FR8221934A FR2538404B1 (de) 1982-12-28 1982-12-28
FR8319506 1983-12-06
FR8319506A FR2555993B1 (fr) 1983-12-06 1983-12-06 Chitosanes 6-sulfates et procede pour leur preparation

Publications (2)

Publication Number Publication Date
EP0116251A1 true EP0116251A1 (de) 1984-08-22
EP0116251B1 EP0116251B1 (de) 1987-12-16

Family

ID=26223212

Family Applications (1)

Application Number Title Priority Date Filing Date
EP83402534A Expired EP0116251B1 (de) 1982-12-28 1983-12-26 Verfahren zum Depolymerisieren und Sulfatieren von Polysacchariden

Country Status (9)

Country Link
EP (1) EP0116251B1 (de)
KR (1) KR920003692B1 (de)
AU (1) AU563377B2 (de)
CA (1) CA1218986A (de)
DE (1) DE3374935D1 (de)
DK (1) DK598083A (de)
IE (1) IE56783B1 (de)
IL (1) IL70511A (de)
NZ (1) NZ206698A (de)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0116801A1 (de) 1982-12-28 1984-08-29 SCLAVO S.p.A. Depolymerisiertes und supersulfatiertes Heparin, Verfahren zu dessen Herstellung und dieses enthaltende pharmazeutische Zusammensetzungen
EP0148057A2 (de) * 1983-12-06 1985-07-10 Sanofi Chitosan-6-sulfat und Verfahren zu dessen Herstellung
EP0166324A2 (de) * 1984-06-16 1986-01-02 B. Braun-SSC AG Verfahren zur selektiven Fällung von Lipoproteinen mit niedriger Dichte
EP0322659A1 (de) * 1987-12-24 1989-07-05 BASF Aktiengesellschaft Verwendung von polysulfatierten Heparinen
EP0356435A1 (de) * 1987-03-19 1990-03-07 Arthropharm Pty Ltd Antientzündungsmittel und zusammensetzungen.
US5013724A (en) * 1985-07-12 1991-05-07 Sanofi Societe Anonyme Process for the sulfation of glycosaminoglycans, the sulfated glycosaminoglycans and their biological applications
US5164378A (en) * 1989-11-24 1992-11-17 Iketon Farmaceutici, S.R.L. Supersulfated heparins
WO1997012621A1 (en) * 1995-09-29 1997-04-10 Novadex Pharmaceuticals Limited Cellulose sulfate for use as antimicrobial and contraceptive agent
US7078392B2 (en) 2000-06-30 2006-07-18 Polydex Pharmaceuticals Limited Cellulose sulfate and other sulfated polysaccharides to prevent and treat papilloma virus infection and other infections
WO2009013162A1 (en) * 2007-07-23 2009-01-29 Istituto Scientifico Di Chimica E Biochimica 'g. Ronzoni' Process for the preparation of heparanase-inhibiting sulfated hyaluronates and products obtained thereby

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2623396B1 (fr) * 1987-11-25 1990-03-30 Sanofi Sa Utilisation de l'ademetionine contre le vieillissement de la peau

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2025073A (en) * 1934-01-09 1935-12-24 Du Pont Cellulose derivative and method of making the same
FR1093999A (fr) * 1952-10-15 1955-05-11 Upjohn Co Procédé de préparation de chitosane sulfaté
US2755275A (en) * 1952-08-29 1956-07-17 Abbott Lab Process for sulfating chitin
US3454560A (en) * 1966-03-01 1969-07-08 Seikagaku Kogyo Co Ltd Process for the production of chondroitin polysulfate

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2025073A (en) * 1934-01-09 1935-12-24 Du Pont Cellulose derivative and method of making the same
US2755275A (en) * 1952-08-29 1956-07-17 Abbott Lab Process for sulfating chitin
FR1093999A (fr) * 1952-10-15 1955-05-11 Upjohn Co Procédé de préparation de chitosane sulfaté
US3454560A (en) * 1966-03-01 1969-07-08 Seikagaku Kogyo Co Ltd Process for the production of chondroitin polysulfate
US3498972A (en) * 1966-03-01 1970-03-03 Kinzo Nagasawa Process of manufacturing dextran sulfate

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0116801A1 (de) 1982-12-28 1984-08-29 SCLAVO S.p.A. Depolymerisiertes und supersulfatiertes Heparin, Verfahren zu dessen Herstellung und dieses enthaltende pharmazeutische Zusammensetzungen
EP0148057A2 (de) * 1983-12-06 1985-07-10 Sanofi Chitosan-6-sulfat und Verfahren zu dessen Herstellung
EP0148057A3 (en) * 1983-12-06 1986-03-26 Anic S.P.A. Chitosan-6-sulphate and process for its preparation
EP0166324A2 (de) * 1984-06-16 1986-01-02 B. Braun-SSC AG Verfahren zur selektiven Fällung von Lipoproteinen mit niedriger Dichte
EP0166324A3 (de) * 1984-06-16 1988-01-13 B. Braun-SSC AG Verfahren zur selektiven Fällung von Lipoproteinen mit niedriger Dichte
US5013724A (en) * 1985-07-12 1991-05-07 Sanofi Societe Anonyme Process for the sulfation of glycosaminoglycans, the sulfated glycosaminoglycans and their biological applications
EP0356435A1 (de) * 1987-03-19 1990-03-07 Arthropharm Pty Ltd Antientzündungsmittel und zusammensetzungen.
EP0356435A4 (de) * 1987-03-19 1990-06-26 Arthropharm Pty Ltd Antientzündungsmittel und zusammensetzungen.
EP0322659A1 (de) * 1987-12-24 1989-07-05 BASF Aktiengesellschaft Verwendung von polysulfatierten Heparinen
US5164378A (en) * 1989-11-24 1992-11-17 Iketon Farmaceutici, S.R.L. Supersulfated heparins
WO1997012621A1 (en) * 1995-09-29 1997-04-10 Novadex Pharmaceuticals Limited Cellulose sulfate for use as antimicrobial and contraceptive agent
US6063773A (en) * 1995-09-29 2000-05-16 Polydex Pharmaceuticals Ltd. Cellulose sulfate for use as antimicrobial and contraceptive agent
US7078392B2 (en) 2000-06-30 2006-07-18 Polydex Pharmaceuticals Limited Cellulose sulfate and other sulfated polysaccharides to prevent and treat papilloma virus infection and other infections
US7226914B2 (en) 2000-06-30 2007-06-05 Rush Presbyterian-St. Luke's Medical Center Cellulose sulfate and other sulfated polysaccharides to prevent and treat papilloma virus infection and other infections
WO2009013162A1 (en) * 2007-07-23 2009-01-29 Istituto Scientifico Di Chimica E Biochimica 'g. Ronzoni' Process for the preparation of heparanase-inhibiting sulfated hyaluronates and products obtained thereby
EP2025687A1 (de) * 2007-07-23 2009-02-18 Istituto Scientifico di Chimica E Biochimica "G Ronzoni Herstellungsverfahren für Heparanase-hemmende sulfierte Hyaluronate und dadurch gewonnene Produkte

Also Published As

Publication number Publication date
CA1218986A (en) 1987-03-10
NZ206698A (en) 1986-11-12
KR840006813A (ko) 1984-12-03
IE833063L (en) 1984-06-28
DK598083D0 (da) 1983-12-23
AU2285683A (en) 1984-07-05
KR920003692B1 (ko) 1992-05-09
DE3374935D1 (en) 1988-01-28
IL70511A (en) 1990-01-18
EP0116251B1 (de) 1987-12-16
DK598083A (da) 1984-06-29
IE56783B1 (en) 1991-12-18
IL70511A0 (en) 1984-03-30
AU563377B2 (en) 1987-07-09

Similar Documents

Publication Publication Date Title
EP0116801B1 (de) Depolymerisiertes und supersulfatiertes Heparin, Verfahren zu dessen Herstellung und dieses enthaltende pharmazeutische Zusammensetzungen
EP0287477B1 (de) Heparine mit niedrigem Molekulargewicht und regelmässiger Struktur, ihre Herstellung und biologische Verwendungen
BE1006827A3 (fr) Melanges de polysaccharides de bas poids moleculaires, procede de preparation et utilisation.
EP0214879B1 (de) Verfahren zur Sulphatierung von Glykosaminoglykanen, daraus hergestellte Glykosaminoglykane und ihre biologischen Verwendungen
US4948881A (en) Process for the depolymerization and sulfation of polysaccharides
CA2501546C (fr) Melanges de polysaccharides derives d'heparine, leur preparation et les compositions pharmaceutiques les contenant
FR2482611A1 (fr) Nouveaux polysaccharides sulfates, procedes pour leur preparation et leur utilisation comme medicaments
CA2416673C (fr) Melanges de polysaccharides derives d'heparine, leur preparation et les compositions pharmaceutiques les contenant
DK160764B (da) Analogifremgangsmaade til fremstilling af terapeutisk aktive heparinfragmenter
EP3298047B1 (de) Methode zur herstellung von polysacchariden
EP0116251A1 (de) Verfahren zum Depolymerisieren und Sulfatieren von Polysacchariden
WO2012114349A1 (en) An improved process for the preparation of pentosan polysulfate sodium
EP0146455A2 (de) Derivate von Dextran mit antikoagulierenden und antikomplementären Eigenschaften, ihre Herstellung und ihre biologischen Anwendungen
Chaubet et al. Synthesis and structure—anticoagulant property relationships of functionalized dextrans: CMDBS
FR2772382A1 (fr) Derives de dextrane, leur procede de preparation et leurs applications comme medicaments a action biologique specifique
EP0356275A1 (de) Selektiv O-acylierte Glycosaminoglykane
RU2512768C1 (ru) Способ получения низкомолекулярного гепарина
EP0148057B1 (de) Chitosan-6-sulfat und Verfahren zu dessen Herstellung
AU2002365302B2 (en) Method for sulphonation of compounds comprising free hydroxyl (OH) groups or primary or secondary amines
JPH02145601A (ja) カードランまたはレンチナンの硫酸エステルまたはその塩の製造方法
FR3047179B1 (fr) Ulvanes sulfates et utilisation dans la regulation des troubles de la coagulation
JP2521083B2 (ja) ヘパリノイド活性を有する多糖体及びその製造方法並びにそれを含有する抗血液凝固剤
RU2034851C1 (ru) Способ получения биологически активного сернокислого эфира хитозана
JPS59138201A (ja) 新規な硫酸化アミノ多糖及びその製造法
BE495863A (de)

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

17P Request for examination filed

Effective date: 19850211

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: SANOFI, SOCIETE ANONYME

Owner name: SCLAVO S.P.A.

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

REF Corresponds to:

Ref document number: 31419

Country of ref document: AT

Date of ref document: 19880115

Kind code of ref document: T

BECN Be: change of holder's name

Effective date: 19871216

REF Corresponds to:

Ref document number: 3374935

Country of ref document: DE

Date of ref document: 19880128

ITF It: translation for a ep patent filed

Owner name: JACOBACCI & PERANI S.P.A.

RAP2 Party data changed (patent owner data changed or rights of a patent transferred)

Owner name: SANOFI, SOCIETE ANONYME

REG Reference to a national code

Ref country code: CH

Ref legal event code: PUE

Owner name: SANOFI, SOCIETE ANONYME

GBT Gb: translation of ep patent filed (gb section 77(6)(a)/1977)
REG Reference to a national code

Ref country code: GB

Ref legal event code: 732

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: AT

Payment date: 19921126

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 19921218

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 19921221

Year of fee payment: 10

Ref country code: CH

Payment date: 19921221

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 19921222

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 19921229

Year of fee payment: 10

ITTA It: last paid annual fee
PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 19921231

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 19930115

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: LU

Payment date: 19930120

Year of fee payment: 10

EPTA Lu: last paid annual fee
PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 19931226

Ref country code: GB

Effective date: 19931226

Ref country code: AT

Effective date: 19931226

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SE

Effective date: 19931227

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LI

Effective date: 19931231

Ref country code: CH

Effective date: 19931231

Ref country code: BE

Effective date: 19931231

BERE Be: lapsed

Owner name: S.A. SANOFI

Effective date: 19931231

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Effective date: 19940701

NLV4 Nl: lapsed or anulled due to non-payment of the annual fee
GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 19931226

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Effective date: 19940831

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DE

Effective date: 19940901

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST

EUG Se: european patent has lapsed

Ref document number: 83402534.8

Effective date: 19940710